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1.
Sensors (Basel) ; 23(23)2023 Nov 21.
Article in English | MEDLINE | ID: mdl-38067671

ABSTRACT

This article provides a comprehensive analysis of the feature extraction methods applied to vibro-acoustic signals (VA signals) in the context of robot-assisted interventions. The primary objective is to extract valuable information from these signals to understand tissue behaviour better and build upon prior research. This study is divided into three key stages: feature extraction using the Cepstrum Transform (CT), Mel-Frequency Cepstral Coefficients (MFCCs), and Fast Chirplet Transform (FCT); dimensionality reduction employing techniques such as Principal Component Analysis (PCA), t-Distributed Stochastic Neighbour Embedding (t-SNE), and Uniform Manifold Approximation and Projection (UMAP); and, finally, classification using a nearest neighbours classifier. The results demonstrate that using feature extraction techniques, especially the combination of CT and MFCC with dimensionality reduction algorithms, yields highly efficient outcomes. The classification metrics (Accuracy, Recall, and F1-score) approach 99%, and the clustering metric is 0.61. The performance of the CT-UMAP combination stands out in the evaluation metrics.


Subject(s)
Robotics , Algorithms , Acoustics , Cluster Analysis , Principal Component Analysis
2.
Cell Tissue Res ; 381(1): 141-161, 2020 Jul.
Article in English | MEDLINE | ID: mdl-32065263

ABSTRACT

Foetal onset hydrocephalus is a disease starting early in embryonic life; in many cases it results from a cell junction pathology of neural stem (NSC) and neural progenitor (NPC) cells forming the ventricular zone (VZ) and sub-ventricular zone (SVZ) of the developing brain. This pathology results in disassembling of VZ and loss of NSC/NPC, a phenomenon known as VZ disruption. At the cerebral aqueduct, VZ disruption triggers hydrocephalus while in the telencephalon, it results in abnormal neurogenesis. This may explain why derivative surgery does not cure hydrocephalus. NSC grafting appears as a therapeutic opportunity. The present investigation was designed to find out whether this is a likely possibility. HTx rats develop hereditary hydrocephalus; 30-40% of newborns are hydrocephalic (hyHTx) while their littermates are not (nHTx). NSC/NPC from the VZ/SVZ of nHTx rats were cultured into neurospheres that were then grafted into a lateral ventricle of 1-, 2- or 7-day-old hyHTx. Once in the cerebrospinal fluid, neurospheres disassembled and the freed NSC homed at the areas of VZ disruption. A population of homed cells generated new multiciliated ependyma at the sites where the ependyma was missing due to the inherited pathology. Another population of NSC homed at the disrupted VZ differentiated into ßIII-tubulin+ spherical cells likely corresponding to neuroblasts that progressed into the parenchyma. The final fate of these cells could not be established due to the protocol used to label the grafted cells. The functional outcomes of NSC grafting in hydrocephalus remain open. The present study establishes an experimental paradigm of NSC/NPC therapy of foetal onset hydrocephalus, at the etiologic level that needs to be further explored with more analytical methodologies.


Subject(s)
Hydrocephalus/therapy , Neural Stem Cells/transplantation , Animals , Cell Differentiation , Cell Proliferation , Neurogenesis , Rats
3.
Polymers (Basel) ; 11(10)2019 Oct 08.
Article in English | MEDLINE | ID: mdl-31597231

ABSTRACT

In this work, we design and produce micron-sized fiber mats by blending poly(ε-caprolactone) (PCL) with small amounts of block copolymers poly(ethylene oxide)m-block-poly(ε-caprolactone)n (PEOm-b-PCLn) using electrospinning. Three different PEOm-b-PCLn block copolymers, with different molecular weights of PEO and PCL, were synthesized by ring opening polymerization of ε-caprolactone using PEO as initiator and stannous octoate as catalyst. The polymer blends were prepared by homogenous solvent mixing using dichloromethane for further electrospinning procedures. After electrospinning, it was found that the addition to PCL of the different block copolymers produced micron-fibers with smaller width, equal or higher hydrophilicity, lower Young modulus, and rougher surfaces, as compared with micron-fibers obtained only with PCL. Neural stem progenitor cells (NSPC), isolated from rat brains and grown as neurospheres, were cultured on the fibrous materials. Immunofluorescence assays showed that the NSPC are able to survive and even differentiate into astrocytes and neurons on the synthetic fibrous materials without any growth factor and using the fibers as guidance. Disassembling of the cells from the NSPC and acquisition of cell specific molecular markers and morphology progressed faster in the presence of the block copolymers, which suggests the role of the hydrophilic character and porous topology of the fiber mats.

4.
J Neuroendocrinol ; 31(3): e12690, 2019 03.
Article in English | MEDLINE | ID: mdl-30697830

ABSTRACT

Tanycytes are located at the base of the brain and retain characteristics from their developmental origins, such as radial glial cells, throughout their life span. With transport mechanisms and modulation of tight junction proteins, tanycytes form a bridge connecting the cerebrospinal fluid with the external limiting basement membrane. They also retain the powers of self-renewal and can differentiate to generate neurones and glia. Similar to radial glia, they are a heterogeneous family with distinct phenotypes. Although the four subtypes so far distinguished display distinct characteristics, further research is likely to reveal new subtypes. In this review, we have re-visited the work of the pioneers in the field, revealing forgotten work that is waiting to inspire new research with today's cutting-edge technologies. We have conducted a systematic ultrastructural study of α-tanycytes that resulted in a wealth of new information, generating numerous questions for future study. We also consider median eminence pituicytes, a closely-related cell type to tanycytes, and attempt to relate pituicyte fine morphology to molecular and functional mechanism. Our rationale was that future research should be guided by a better understanding of the early pioneering work in the field, which may currently be overlooked when interpreting newer data or designing new investigations.


Subject(s)
Brain/cytology , Brain/physiology , Ependymoglial Cells/physiology , Ependymoglial Cells/ultrastructure , Animals , Blood-Brain Barrier/cytology , Blood-Brain Barrier/physiology , Cerebrospinal Fluid/cytology , Humans , Median Eminence/ultrastructure , Third Ventricle/ultrastructure
5.
Cell Tissue Res ; 373(2): 421-438, 2018 Aug.
Article in English | MEDLINE | ID: mdl-29651556

ABSTRACT

Fetal onset hydrocephalus and abnormal neurogenesis are two inseparable phenomena turned on by a cell junction pathology first affecting neural stem/progenitor cells (NSPCs) and later the multiciliated ependyma. The neurological impairment of children born with hydrocephalus is not reverted by derivative surgery. NSPCs and neurosphere (NE) grafting into the cerebrospinal fluid (CSF) of hydrocephalic fetuses thus appears as a promising therapeutic procedure. There is little information about the cell lineages actually forming the NE as they grow throughout their days in vitro (DIV). Furthermore, there is no information on how good a host the CSF is for grafted NE. Here, we use the HTx rat, a model with hereditary hydrocephalus, with the mutation expressed in about 30% of the litter (hyHTx), while the littermates develop normally (nHTx). The investigation was designed (i) to establish the nature of the cells forming 4 and 6-DIV NE grown from NSPCs collected from PN1/nHTx rats and (ii) to study the effects on these NEs of CSF collected from nHTx and hyHTx. Immunofluorescence analyses showed that 90% of cells forming 4-DIV NEs were non-committed multipotential NSPCs, while in 6-DIV NE, 40% of the NSPCs were already committed into neuronal, glial and ependymal lineages. Six-DIV NE further cultured for 3 weeks in the presence of fetal bovine serum, CSF from nHTx or CSF from hyHTx, differentiated into neurons, astrocytes and ßIV-tubulin+ multiciliated ependymal cells that were joined together by adherent junctions and displayed synchronized cilia beating. This supports the possibility that ependymal cells are born from subpopulations of NSC with their own time table of differentiation. As a whole, the findings indicate that the CSF is a supportive medium to host NE and that NE grafted into the CSF have the potential to produce neurons, glia and ependyma.


Subject(s)
Astrocytes/cytology , Cerebrospinal Fluid/physiology , Ependyma/cytology , Ependymoglial Cells/cytology , Hydrocephalus/pathology , Neural Stem Cells/metabolism , Animals , Cell Differentiation , Cell Proliferation , Cilia/metabolism , Disease Models, Animal , Humans , Multipotent Stem Cells/cytology , Multipotent Stem Cells/metabolism , Neural Stem Cells/cytology , Neurons/cytology , Rats
6.
Fluids Barriers CNS ; 11: 7, 2014.
Article in English | MEDLINE | ID: mdl-24685106

ABSTRACT

I was honored to be awarded the Casey Holter Essay Prize in 2013 by the Society for Research into Hydrocephalus and Spina Bifida. The purpose of the prize is to encourage original thinking in a way to improve the care of individuals with spina bifida and hydrocephalus. Having kept this purpose in mind, I have chosen the title: Neural stem cells, are they the hope of a better life for patients with fetal-onset hydrocephalus? The aim is to review and discuss some of the most recent and relevant findings regarding mechanisms leading to both hydrocephalus and abnormal neuro/gliogenesis. By looking at these outcome studies, it is hoped that we will recognize the potential use of neural stem cells in the treatment of hydrocephalus, and so prevent the disease or diminish/repair the associated brain damage.

7.
Cell Tissue Res ; 352(3): 707-25, 2013 Jun.
Article in English | MEDLINE | ID: mdl-23640132

ABSTRACT

The present investigation was designed to clarify the role of the subcommissural organ (SCO) in the pathogenesis of hydrocephalus occurring in the HTx rat. The brains of non-affected and hydrocephalic HTx rats from embryonic day 15 (E15) to postnatal day 10 (PN10) were processed for electron microscopy, lectin binding and immunocytochemistry by using a series of antibodies. Cerebrospinal fluid (CSF) samples of non-affected and hydrocephalic HTx rats were collected at PN1, PN7 and PN30 and analysed by one- and two-dimensional electrophoresis, immunoblotting and nanoLC-ESI-MS/MS. A distinct malformation of the SCO is present as early as E15. Since stenosis of the Sylvius aqueduct (SA) occurs at E18 and dilation of the lateral ventricles starts at E19, the malformation of the SCO clearly precedes the onset of hydrocephalus. In the affected rats, the cephalic and caudal thirds of the SCO showed high secretory activity with all methods used, whereas the middle third showed no signs of secretion. At E18, the middle non-secretory third of the SCO progressively fused with the ventral wall of SA, resulting in marked aqueduct stenosis and severe hydrocephalus. The abnormal development of the SCO resulted in the permanent absence of Reissner's fibre (RF) and led to changes in the protein composition of the CSF. Since the SCO is the source of a large mass of sialilated glycoproteins that form the RF and of those that remain CSF-soluble, we hypothesize that the absence of this large mass of negatively charged molecules from the SA domain results in SA stenosis and impairs the bulk flow of CSF through the aqueduct.


Subject(s)
Hydrocephalus/etiology , Hydrocephalus/pathology , Subcommissural Organ/pathology , Amino Acid Sequence , Animals , Cell Differentiation , Cerebral Aqueduct/metabolism , Cerebral Aqueduct/pathology , Cerebral Aqueduct/ultrastructure , Constriction, Pathologic , Embryo, Mammalian/pathology , Fetus/pathology , Hydrocephalus/cerebrospinal fluid , Molecular Sequence Data , Prealbumin/cerebrospinal fluid , Prealbumin/chemistry , Rats , Subcommissural Organ/metabolism , Subcommissural Organ/ultrastructure
8.
Reprod Fertil Dev ; 23(3): 417-23, 2011.
Article in English | MEDLINE | ID: mdl-21426859

ABSTRACT

In the present study, we analysed the molecular mechanism(s) by which melatonin directly affects ovarian function in the mare. In Experiment 1, follicles and corpora lutea (CL) were collected from slaughterhouse ovaries and analysed for melatonin (MT(1)) receptor mRNA and protein. In Experiment 2, CL were collected from slaughterhouse ovaries and cultured in Dulbecco's modified Eagle's medium-F12 medium (control medium) supplemented with 50 ng mL(-1) equine chorionic gonadotrophin (eCG), 1 nM-1 µM melatonin, 1 µM forskolin or 1 µM luzindole. Explants were cultured for 3 h in the presence of these drugs. Conditioned media were analysed for progesterone production; luteal cells were analysed for cholesterol side-chain cleavage enzyme (P450scc), a steroidogenic enzyme that converts cholesterol into pregnenolone. Both MT(1) receptor mRNA and protein were expressed in follicles and CL. Melatonin inhibited both the eCG- and forskolin-stimulated production of progesterone, as well as the forskolin-stimulated expression of P450scc, in equine luteal cells and the effect was dose-dependent. The inhibitory effect of melatonin was blocked by luzindole, a non-selective melatonin MT(1) and MT(2) receptor antagonist. The data support the presence of functional melatonin receptors in luteal cells and a regulatory role for melatonin in the endocrine function of the equine CL.


Subject(s)
Horses/physiology , Luteal Cells/drug effects , Luteal Cells/metabolism , Melatonin/pharmacology , Ovarian Follicle/metabolism , Progesterone/metabolism , Receptor, Melatonin, MT1/biosynthesis , Animals , Blotting, Western/veterinary , Cholesterol Side-Chain Cleavage Enzyme/biosynthesis , Cholesterol Side-Chain Cleavage Enzyme/metabolism , Chorionic Gonadotropin/pharmacology , Colforsin/pharmacology , Female , Ovarian Follicle/cytology , Ovarian Follicle/drug effects , RNA, Messenger/biosynthesis , RNA, Messenger/genetics , Receptor, Melatonin, MT1/antagonists & inhibitors , Receptor, Melatonin, MT1/genetics , Reverse Transcriptase Polymerase Chain Reaction/veterinary , Statistics, Nonparametric , Tryptamines/pharmacology
9.
Acta Neuropathol ; 121(6): 721-35, 2011 Jun.
Article in English | MEDLINE | ID: mdl-21311902

ABSTRACT

A heterogeneous population of ependymal cells lines the brain ventricles. The evidence about the origin and birth dates of these cell populations is scarce. Furthermore, the possibility that mature ependymal cells are born (ependymogenesis) or self-renewed (ependymal proliferation) postnatally is controversial. The present study was designed to investigate both phenomena in wild-type (wt) and hydrocephalic α-SNAP mutant (hyh) mice at different postnatal stages. In wt mice, proliferating cells in the ventricular zone (VZ) were only found in two distinct regions: the dorsal walls of the third ventricle and Sylvian aqueduct (SA). Most proliferating cells were monociliated and nestin+, likely corresponding to radial glial cells. Postnatal cumulative BrdU-labeling showed that most daughter cells remained in the VZ of both regions and they lost nestin-immunoreactivity. Furthermore, some labeled cells became multiciliated and GLUT-1+, indicating they were ependymal cells born postnatally. Postnatal pulse BrdU-labeling and Ki-67 immunostaining further demonstrated the presence of cycling multiciliated ependymal cells. In hydrocephalic mutants, the dorsal walls of the third ventricle and SA expanded enormously and showed neither ependymal disruption nor ventriculostomies. This phenomenon was sustained by an increased ependymogenesis. Consequently, in addition to the physical and geometrical mechanisms traditionally explaining ventricular enlargement in fetal-onset hydrocephalus, we propose that postnatal ependymogenesis could also play a role. Furthermore, as generation of new ependymal cells during postnatal stages was observed in distinct regions of the ventricular walls, such as the roof of the third ventricle, it may be a key mechanism involved in the development of human type 1 interhemispheric cysts.


Subject(s)
Brain/pathology , Ependyma/growth & development , Hydrocephalus/pathology , Third Ventricle/physiopathology , Age Factors , Animals , Animals, Newborn , Bromodeoxyuridine/metabolism , Cell Count , Cell Proliferation , Disease Models, Animal , Ependyma/ultrastructure , Gene Expression Regulation, Developmental/physiology , Glial Fibrillary Acidic Protein/metabolism , Glucose Transporter Type 1/metabolism , Mice , Mice, Neurologic Mutants , Microscopy, Electron, Scanning , Proliferating Cell Nuclear Antigen/metabolism , Third Ventricle/cytology , Tubulin/metabolism
10.
Peptides ; 31(4): 757-76, 2010 Apr.
Article in English | MEDLINE | ID: mdl-20093161

ABSTRACT

The blood-brain barrier (BBB) is a single uninterrupted barrier that in the brain capillaries is located at the endothelial cells and in the circumventricular organs, such as the choroid plexuses (CP) and median eminence (ME), is displaced to specialized ependymal cells. How do hypothalamic hormones reach the portal circulation without making the BBB leaky? The ME milieu is open to the portal vessels, while it is closed to the cerebrospinal fluid (CSF) and to the arcuate nucleus. The cell body and most of the axons of neurons projecting to the ME are localized in areas protected by the BBB, while the axon terminals are localized in the BBB-free area of the ME. This design implies a complex organization of the intercellular space of the median basal hypothalamus. The privacy of the ME milieu implies that those neurons projecting to this area would not be under the influence of compounds leaking from the portal capillaries, unless receptors for such compounds are located at the axon terminal. Amazingly, the arcuate nucleus also has its private milieu that is closed to all adjacent neural structures and open to the infundibular recess. The absence of multiciliated cells in this recess should result in a slow CSF flow at this level. This whole arrangement should facilitate the arrival of CSF signal to the arcuate nucleus. This review will show how peripheral hormones can reach hypothalamic targets without making the BBB leaky.


Subject(s)
Arcuate Nucleus of Hypothalamus/anatomy & histology , Blood-Brain Barrier/physiology , Cerebrospinal Fluid/metabolism , Hypothalamus/anatomy & histology , Median Eminence/anatomy & histology , Animals , Arcuate Nucleus of Hypothalamus/metabolism , Blood-Brain Barrier/ultrastructure , Hypothalamus/metabolism , Median Eminence/metabolism , Neurons/cytology , Neurons/physiology , Pituitary Hormones/metabolism
11.
Cell Tissue Res ; 339(2): 359-81, 2010 Feb.
Article in English | MEDLINE | ID: mdl-19937347

ABSTRACT

The pars tuberalis (PT) is the only pituitary region in close contact with the medial-basal hypothalamus and bathed by cerebrospinal fluid (CSF). Although PT has long been recognized as an endocrine gland, certain aspects of its structure remain obscure. The present investigation has been designed to gain information concerning (1) the cellular organization of PT, (2) the PT/median eminence spatial relationship and (3) the exposure of various cell compartments of PT to CSF. Non-endocrine cells (S100-reactive) appear as the organizer of the PT architecture. The apical poles of these cells line large cistern-like cavities and the processes of these cells establish a close spatial relationship with PT-specific secretory cells, portal capillaries and tanycytes. The cisterns are also endowed with clusters of ciliated cells and with a highly electron-dense and PAS-reactive content. The unique spatial organization of endocrine and non-endocrine cells of the PT supports a functional relationship between both cell populations. PT endocrine cells display a hallmark of PT-specific cells, namely, the paranuclear spot, which is a complex structure involving the Golgi apparatus, a large pool of immature secretory granules and a centriole from which originates a single 9+0 cilium projecting to the intercellular channels. Horseradish peroxidase (HRP) injected into the CSF readily reaches the intercellular channels of PT and the inner channel of the single cilium and is incorporated by the endocytic machinery of the secretory cells. The PT endocrine cells, through their single 9+0 cilium, may act as sensors of the CSF. HRP also reaches the lumen of the cisterns, indicating that this PT compartment is also exposed to CSF. PT endocrine cells establish direct cell-to-cell contacts with hypothalamic beta(1) tanycytes, suggesting a second means of brain-PT communication.


Subject(s)
Cerebrospinal Fluid , Ependyma/cytology , Median Eminence/cytology , Pituitary Gland, Anterior/cytology , Animals , Capillaries , Centrioles/ultrastructure , Cilia/ultrastructure , Endocrine Cells/metabolism , Endocrine Cells/ultrastructure , Endocytosis , Extracellular Space , Golgi Apparatus/metabolism , Golgi Apparatus/ultrastructure , Hypothalamus, Middle/cytology , Pituitary Gland, Anterior/metabolism , Rats , S100 Proteins/metabolism , Secretory Vesicles/ultrastructure , Third Ventricle/cytology
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